Introduction to Lab Ex. 23: Hemagglutination

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Introduction to Lab Ex. 23 Hemagglutination
Complement Mediated Lysis
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Introduction to Lab Ex.23 Hemagglutination Co
mplement Mediated Lysis Agglutination is the
clumping of particulate antigens through their
binding with specific antibodies. Hemagglutinat
ion is the clumping of red blood cells through
their binding with specific antibodies. Thus
hemagglutination is used as a observable reaction
between specific antibodies with red blood
cells. Complement is a series of proteins found
in the serum with non-specific immune function
resulting in the lysis of target cells.
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Properties of complement include - non-specific
immune function - inactivated by heat (560C for
30mins) - consists of 9 different proteins in a
cascade - activated by specific antigen
antibody complex formation (classical) certain
chemicals endotoxin (alternate)
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Hemagglutination

• Hemagglutination involves agglutination of RBCs.
• Viral hemagglutination inhibition tests for
  antibodies by the antibodies' ability to prevent
  viruses from agglutinating RBCs.

Figure 18.7
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The Complement System

• Serum proteins activated in a cascade.

Figure 16.10
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Effects of Complement Activation

• Opsonization or immune adherence enhanced
  phagocytosis
• Membrane attack complex cytolysis
• Attract phagocytes

Figure 16.11
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Effects of Complement Activation
Figure 16.12
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Experiment Agantigen Ab antibody C
complement Premise Antigen used is sheep RBC
(sRBC) Antibody used is anti-sRBC - antibody to
sRBC (from immunized rabbit blood) Normal
serum is from normal rabbit (Ab absent) Immune
serum is from rabbit immunized with sRBC (Ab
present) Normal serum (Ab absent C
present) Immune serum (Ab present C
present) Normal serum heated (Ab absent C
absent) Immune serum heated (Ab present C
absent)
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Legend Ag sRBC Ab antibody to sRBC C
complement Tube 1 sRBC saline (Ag)
No reaction
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Legend Ag sRBC Ab antibody to sRBC C
complement
Tube 2 sRBC Normal serum (Ag) (-AbC)
No reaction
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Legend Ag sRBC Ab antibody to sRBC C
complement
Tube 3 sRBC Immune serum (Ag) (AbC)
Hemolysis
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Legend Ag sRBC Ab antibody to sRBC C
complement
Tube 4 sRBC Normal serum (heated)
(Ag) (-Ab-C)
No reaction
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Legend Ag sRBC Ab antibody to sRBC C
complement
Tube 5 sRBC Immune serum (heated)
(Ag) (Ab-C)
Hemagglutination
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Legend Ag sRBC Ab antibody to sRBC C
complement
Tube 6 sRBC Normal serum Heated Immune serum
(Ag) (-AbC) (Ab-C)
Hemolysis
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Legend Ag sRBC Ab antibody to sRBC C
complement Tube 1 sRBC saline No
reaction Tube 2 sRBC Normal serum No
reaction Tube 3 sRBC Immune serum
Hemolysis Tube 4 sRBC heated normal serum
No reaction Tube 5 sRBC heated Immune
serum Hemagglutination Tube 6 sRBC Normal
serum heated Immune serum hemoly
sis
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Legend Ag sRBC Ab antibody to sRBC C
complement Tube 1 sRBC saline No
reaction Tube 2 sRBC Normal serum No
reaction Tube 3 sRBC Immune serum
Hemolysis Tube 4 sRBC heated normal serum
No reaction Tube 5 sRBC heated Immune serum
Hemagglutination Tube 6 sRBC Normal serum
heated Immune serum hemolysis